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Literature summary for 1.1.1.337 extracted from

  • Feil, I.K.; Hendle, J.; Schomburg, D.
    Modified substrate specificity of L-hydroxyisocaproate dehydrogenase derived from structure-based protein engineering (1997), Protein Eng., 10, 255-262.
    View publication on PubMed
Search for more literature for 1.1.1.337

Protein Variants

Protein Variants Comment Organism
F236S phenylpyruvate displays the largest kcat of the tested substrates. All kcat values, with the exception of phenylpyruvate, are reduced, but the relative acceptance of phenylglyoxylate is greatly increased Weissella confusa
F236V decrease in the second-order rate constants for all tested substrates. The kcat values for the smaller substrates decrease drastically. Phenylpyruvate is the favourite substrate (2-oxocaproate is the favourite substrate of the wild-type enzyme) Weissella confusa
G234V/G235D the second-order rate constant decreased for most substrates with the exception of pyruvate, which reacts 2.5times faster in the mutant enzyme than in the wild-type enzyme. The catalysis of 2-oxoisocaproate is unaffected Weissella confusa
L239A shift of enzyme specificity towards the substrates branched at C3 corresponding to an increase in the turnover numbers for 2-oxoisocaproate Weissella confusa
L239F slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values Weissella confusa
L239M KM values of all substrates of the enzyme variant are very similar to those of the wild-type enzyme, large improvement in the kcat values of the C4-branched substrates 2-oxoisocaproate and phenylpyruvate, decrease in the turnover number of 2-oxocaproate (the substrate favoured by the wild-type enzyme). Whereas in the wild-type enzyme the second-order rate constant for 2-oxoisocaproate is only 1% of that for the unbranched substrate, in the mutant enzyme the rate constant of 2-oxocaproate is only 18% of that for 2-oxoisocaproate Weissella confusa
L239M/T245A the catalytic rates are reduced by several orders of magnitude and the KM values shows at least a 100fold increase for most substrates (with the exception of phenylglyoxylate). With respect to L239M the substrate specificity shifts towards keto-tert-leucine and with respect to T245A 2-oxoisocaproate and phenylpyruvate are more favoured Weissella confusa
L239W slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values Weissella confusa
T245A the catalytic rates are reduced by several orders of magnitude, relative shift of substrate specificity for keto-tert-leucine of more than 17 000 compared with the 2-oxocaproate (kcat/KM) Weissella confusa

Organism

Organism UniProt Comment Textmining
Weissella confusa P14295 i.e. Lactobacillus confusus
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxocaproate + NADH + H+ 2-oxocaproate is the best substrate for the wild-type enzyme Weissella confusa 2-hydroxycaproate + NAD+
-
 r
2-oxoisocaproate + NADH + H+
-
 Weissella confusa 2-hydroxyisocaproate + NAD+
-
 r
additional information broad substrate specificity, utilizes a wide range of 2-oxo acids branched at the C4 atom Weissella confusa ?
-
 ?
oxo-tert-leucine + NADH + H+
-
 Weissella confusa ? + NAD+
-
 r
phenylglyoxylate + NADH + H+
-
 Weissella confusa ? + NAD+
-
 r
phenylpyruvate + NADH + H+
-
 Weissella confusa 3-phenyllactate + NAD+
-
 r
pyruvate + NADH + H+
-
 Weissella confusa L-lactate + NAD+
-
 r

Synonyms

Synonyms Comment Organism
L-HicDH
-
 Weissella confusa
L-hydroxyisocaproate dehydrogenase
-
 Weissella confusa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.07
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
0.075
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
0.1
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
0.1
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
0.2
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
0.2
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
0.2
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
0.3
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
0.4
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
0.9
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.3
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
2 8 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
6.58
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
9.5
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
20
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
20
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
22
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
25
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
27
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
32
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
51
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
52
-
 oxo-tert-leucine pH 7.0, 30°C, wild-type enzyme Weissella confusa
90
-
 pyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
92
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
100
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
150
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
210
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
320
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
340
-
 pyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
350
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
420
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
520
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
920
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
2100
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
2400
-
 2-oxoisocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa
2400
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
2500
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
2500
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
3400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
3500
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
4100
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
4400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
5600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
6400
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
6900
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
7300
-
 phenylpyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
7600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
7900
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
11000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
13000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
15000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
18000
-
 phenylglyoxylate pH 7.0, 30°C, wild-type enzyme Weissella confusa
360000
-
 2-oxocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00526
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
0.01
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
0.012
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
0.0465
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
0.0469
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
0.0684
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
0.07
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
0.357
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
0.411
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
0.654
-
 pyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.08
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.82
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
2.97
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
3.09
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
3.84
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
5.88
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
6.06
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
6.13
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
10
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
13
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
15
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
16
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
22.5
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
25.7
-
 pyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
32.8
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
66.2
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
117
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
133
-
 oxo-tert-leucine pH 7.0, 30°C, wild-type enzyme Weissella confusa
158
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
677
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
941
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1460
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1530
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1880
-
 phenylglyoxylate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1920
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
2190
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
2420
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
2620
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
2670
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
2880
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
5060
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
7330
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
10000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
28700
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
35800
-
 2-oxoisocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa
37600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
39100
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
49200
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
76000
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
203000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
224000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
281000
-
 phenylpyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
3270000
-
 2-oxocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa